摘要
目的利用SELEX技术筛选HIV-P24的核酸适配体,为艾滋病的诊断和治疗奠定基础。方法以重组P24为筛选靶,用SELEX技术从随机寡核苷酸库中筛选与HIV-P24结合的寡核苷酸,利用凝胶阻滞实验鉴定第12轮筛选到的寡核苷酸与HIV-P24的结合,再用Dot-blot法筛选出与HIV-P24结合的核酸适配体,并检测核酸适配体识别HIV-P24的特异性。结果 Dot-blot筛选到5条与HIV-P24有较强结合能力的核酸适配体,且均为不同的序列。特异性检测显示,18和26号配体只与HIV-P24特异性结合,而与人血清白蛋白、牛血清白蛋白和脱脂奶粉均无明显结合。结论成功筛选到2条特异结合HIV-P24的核酸适配体,为其应用于艾滋病诊断和治疗提供了实验基础。
Objective To screen aptamers to HIV-P24 through AIDS. Methods With recombinant P24 for the screening SELEX technique for the diagnosis and therapy of target, oligonucleotides binding to HIV-P24 were screened from a random oligonucleotide library through SELEX technique. The binding capacity between oligonu- cleotides obtained from the 12th round of screening and HIV-P24 was identified via electrophoretic mobility shift as- say （EMSA）. The aptamers strongly binding to HIV-P24 were screened and the recognition specificity of aptamers for HIV-P24 was detected by Dot-blot method. Results Five aptamers with high affinity to HIV-P24 were obtained with different sequences. The binding specificity showed that No. 18 and No. 26 apatmers only bound to HIV-P24, not to human serum albumin, bovine serum albumin and skimmed milk powder. Conclusions Two aptamers spe- cially binding to HIV-P24 were obtained, and thus provides an experimental basis for the diagnosis and treatment of AIDS by utilizing aptamer of HIV-P24.
出处
《基础医学与临床》
CSCD
北大核心
2012年第9期987-991,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(30271219
30960104
81060180)
国家中小企业创新基金(06C26226200591)
甘肃省重大研发项目(GKSOU-B06-10
GKS031030)
